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The minimal promoter region of the dense-core vesicle protein IA-2: transcriptional regulation by CREB

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Abstract

Aims

IA-2 is a transmembrane protein found in the dense-core vesicles (DCV) of neuroendocrine cells and one of the major autoantigens in type 1 diabetes. DCV are involved in the secretion of hormones (e.g., insulin) and neurotransmitters. Stimulation of pancreatic β cells with glucose upregulates the expression of IA-2 and an increase in IA-2 results in an increase in the number of DCV. Little is known, however, about the promoter region of IA-2 or the transcriptional factors that regulate the expression of this gene.

Methods

In the present study, we constructed eight deletion fragments from the upstream region of the IA-2 transcription start site and linked them to a luciferase reporter.

Results

By this approach, we have identified a short bp region (−216 to +115) that has strong promoter activity. We also identified a transcription factor, cAMP responsive element-binding protein (CREB), which binds to two CREB-related binding sites located in this region. The binding of CREB to these sites enhanced IA-2 transcription by more than fivefold. We confirmed these findings by site-directed mutagenesis, chromatin immunoprecipitation assays and RNAi inhibition.

Conclusion

Based on these findings, we conclude that the PKA pathway is a critical, but not the exclusive signaling pathway involved in IA-2 gene expression.

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Acknowledgments

We thank Dr. Raju Gottumukkala for his comments and Charles Wohlenberg for technical help. This research was supported by the Intramural Research Program of the NIH.

Conflict of interest

The authors declare that there is no conflict of interest that would prejudice the impartiality of this scientific work.

Ethical standard

All studies were performed according to NIH guidelines.

Human and animal rights disclosure

This article does not contain any studies with human or animal subjects performed by the any of the authors.

Informed consent disclosure

Informed consent was obtained from all participants prior to inclusion in the study.

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Authors and Affiliations

  1. Experimental Medicine Section, Laboratory of Sensory Biology, National Institute of Dental and Craniofacial Research (NIDCR), National Institutes of Health (NIH), B30/Rm106, Bethesda, MD, 20892, USA

    Tao Cai, Hiroki Hirai, Huanyu Xu & Abner L. Notkins

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  1. Tao Cai
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  2. Hiroki Hirai
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  3. Huanyu Xu
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Correspondence to Abner L. Notkins.

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Cai, T., Hirai, H., Xu, H. et al. The minimal promoter region of the dense-core vesicle protein IA-2: transcriptional regulation by CREB. Acta Diabetol 52, 573–580 (2015). https://doi.org/10.1007/s00592-014-0689-5

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  • DOI: https://doi.org/10.1007/s00592-014-0689-5

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